The DNM3 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population for loss-of-function studies of human dynamin-3 (DNM3). This product is a heterogeneous pool of HAP1 cells with targeted disruptions in DNM3, generated by CRISPR/Cas9 genome editing. As a polyclonal model, it captures diverse editing outcomes, making it suitable for pooled genetic screens, bulk endocytosis assays, and drug response profiling without requiring single-cell clonality. The knockout population can be used directly in functional experiments, providing a cost-effective alternative to monoclonal lines.
HAP1 is a human near-haploid fibroblast-like cell line derived from a male patient with chronic myeloid leukemia (CML). Its near-haploid genome facilitates genetic analysis and CRISPR-based screening, as most genes are single-copy, enabling efficient knockout generation and clear phenotype interpretation. HAP1 cells maintain functional signaling pathways and endocytic machinery, allowing investigation of membrane trafficking, cytoskeletal dynamics, and cancer-relevant processes. This line is widely used in high-throughput genetic screens and functional genomics.
DNM3 encodes a dynamin GTPase mediating membrane fission in clathrin-mediated endocytosis. It oligomerizes at vesicle necks and undergoes GTP hydrolysis-driven conformational changes that sever clathrin-coated vesicles from the plasma membrane. DNM3 also regulates actin reorganization by interacting with cortactin and profilin. Upstream regulators include endophilin, amphiphysin, GSK3??, and CDK5; DNM3 acts downstream on clathrin-coated vesicles, actin filaments, and synaptic vesicles. It binds SH3-domain-containing proteins like Grb2 and endophilin A1?CA3, linking trafficking to signaling. This places DNM3 at the intersection of endocytosis, actin dynamics, and possibly mitochondrial function, impacting synaptic recycling and cell migration.
In the HAP1 context, DNM3 knockout provides a unique system to dissect endocytic pathways in a near-haploid, cancer-derived cell background. Disruption of DNM3 impairs clathrin-mediated endocytosis, enabling quantification of transferrin uptake or receptor trafficking changes. Given HAP1??s leukemic origin, this model facilitates studies on dynamin3-dependent trafficking in cancer cell behaviors such as proliferation, migration, and chemotherapeutic response. The polyclonal population may reveal phenotypic heterogeneity and resistance mechanisms in drug sensitivity screens, enhancing its utility for basic and translational research.
This knockout model is ideal for functional studies of endocytosis and membrane trafficking, with representative assays including western blotting to verify DNM3 loss, transferrin uptake assays for endocytosis efficiency, immunofluorescence staining of clathrin-coated structures, wound healing assays for cell migration, and co-immunoprecipitation to probe DNM3 interactomes. In cancer research, the cells can test drug sensitivities in CML models where DNM3 influences chemotherapeutic responses. The polyclonal population is also suited for high-throughput genetic screens to identify endocytic regulators or drug resistance modulators. For further information, please contact Ascent Research.
